CN105713990A - Wheat molecular marker and application thereof in identifying wheat yield related traits - Google Patents
Wheat molecular marker and application thereof in identifying wheat yield related traits Download PDFInfo
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Abstract
The invention discloses a wheat molecular marker and application thereof in identifying wheat yield related traits. According to the wheat molecular marker, a primer pair P is utilized to perform amplification by using wheat genome DNA (deoxyribonucleic acid) as a template to obtain DNA molecules. The primer pair P is composed of single-stranded DNAs p1 and p2, wherein the p1 is a single-stranded DNA which is specifically combined with the forward 70th site disclosed as Sequence 1 in the wheat genome DNA, and the p2 is a single-stranded DNA which is specifically combined with the reverse 241st site disclosed as Sequence 1 in the wheat genome DNA. The wheat is subjected to genotyping according to the wheat molecular marker to obtain the three homozygous genotypes A1A1, A2A2 and A3A3. The experiment proves that the genotypes A1A1, A2A2 and A3A3 of the wheat are related to wheat yield related traits-number of spikelets per spike, number of ears per plant and weight of thousand grains, and can be used for culturing high-yield wheat species.
Description
Technical field
The present invention relates to wheat molecular marker and the application in identifying wheat yield correlated traits thereof in biological technical field.
Background technology
Semen Tritici aestivi (TriticumaestivumL.) is as one of main cereal crops in the world, and its safety in production is the important leverage of mankind's grain security.And this cost-effective approach of kind that selects and breeds high yielding wheat, mankind's grain-production is had safely particularly important strategic importance.Conventional breeding generally carries out Phenotypic Selection, although also obtaining very big progress, but time and effort consuming；Comparatively speaking, molecular mark strategy is relatively simple easy and can efficiently utilize favorable genes, thus dramatically speeding up breeding process.
Summary of the invention
The technical problem to be solved is how to identify wheat yield correlated traits, such as mass of 1000 kernel, every fringe spikelet number and/or single-strain tassel number.
For solving above-mentioned technical problem, present invention firstly provides wheat molecular marker in the application identified or in auxiliary qualification wheat yield correlated traits；
Described wheat molecular marker, its name is called CAPS-7A, for being template with the genomic DNA of Semen Tritici aestivi, adopt primer pair P to carry out expanding the DNA molecular obtained；By name, described primer pair P is called that the single stranded DNA of p1 and p2 forms, described p1 is and the single stranded DNA of the 70th upstream specific bond of sequence 1 in Wheat volatiles DNA, and described p2 is and the single stranded DNA of the 241st downstream specific bond of sequence 1 in Wheat volatiles DNA.
Concrete, described wheat molecular marker, is following 1 for the 70th-240 corresponding to sequence 1 in Semen Tritici aestivi A genomic DNA), 2) or 3):
1) the 70th-240 of sequence 1；
2) the 70th-240 of sequence 2；
3) the 70th-254 of sequence 3.
Sequence 1 is made up of 294 nucleotide, and sequence 2 is made up of 294 nucleotide, and sequence 3 is made up of 307 nucleotide.Sequence 1 is only different at the 218th from the sequence of sequence 2, and the 218th of sequence 1 is T, and the 218th of sequence 2 is G；Sequence 3 has three places different from sequence 1, and sequence 3 is insert TCTCTCTCTCTCTC at the 70th of sequence 1 with the 71st interdigit, the T of the 218th of sequence 1 the sports G and the C of the 240th of sequence 1 the is lacked the sequence obtained.
In above-mentioned application, described p1 can for the single stranded DNA shown in sequence 4, and described p2 can for the single stranded DNA shown in sequence 5.
For solving above-mentioned technical problem, present invention also offers and identify or assist the primer pair identifying wheat yield correlated traits, described primer pair is described primer pair P.
The mol ratio of two single stranded DNAs of described primer pair P can be 1:1.Two single stranded DNAs of described primer pair P can independent packaging.
For solving above-mentioned technical problem, present invention also offers and identify or assist the reagent set identifying wheat yield correlated traits, described reagent set is made up of described primer pair P and restricted enzyme Nco I.
Wherein, described primer pair P and restricted enzyme Nco I can independent packaging.
For solving above-mentioned technical problem, present invention also offers and identify or assist the method identifying wheat genotypes, described genotype is A1A1 genotype, A2A2 genotype, A3A3 genotype, A1A2 genotype, A1A3 genotype and A2A3 genotype, and described method is following L, M, N or O:
L, include following L1) and L2):
L1) with Wheat volatiles DNA to be measured for template, utilize described primer pair P to carry out pcr amplification and obtain PCR primer；
L2) detecting step L1) sequence of PCR primer that obtains, if described PCR primer contains the DNA fragmentation shown in X1 but do not contain the DNA fragmentation shown in X2 or X3, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X2 and without there being the DNA fragmentation shown in described X1 or described X3, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X3 and without there being the DNA fragmentation shown in described X1 or described X2, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X1 and described X2 and without there being the DNA fragmentation shown in described X3, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X1 and described X3 and without there being the DNA fragmentation shown in described X2, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X2 and described X3 and without there being the DNA fragmentation shown in described X1, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
Described X1 is the 70th-240 of sequence 1；
Described X2 is the 70th-240 of sequence 2；
Described X3 is the 70th-254 of sequence 3；
M, include following M1) and M2):
M1) with Wheat volatiles DNA to be measured for template, utilize described primer pair P to carry out pcr amplification and obtain PCR primer；
M2) detecting step M1) sequence of PCR primer that obtains, if the sequence of described PCR primer is sequence 1, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If the sequence of described PCR primer is sequence 2, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If the sequence of described PCR primer is sequence 3, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If the sequence of described PCR primer is sequence 1 and sequence 2, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If the sequence of described PCR primer is sequence 1 and sequence 3, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If the sequence of described PCR primer is sequence 2 and sequence 3, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
N, include following N1) and N2):
N1) with Wheat volatiles DNA to be measured for template, utilize described primer pair P to carry out pcr amplification and obtain PCR primer, adopt restricted enzyme NcoI to process described PCR primer and obtain digestion products；
N2) detecting step N1) size of digestion products that obtains, if the DNA fragmentation that described digestion products is a 294bp, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If the DNA fragmentation that described digestion products is two size respectively 215bp and 79bp, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If the DNA fragmentation that described digestion products is two size respectively 229bp and 78bp, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If the DNA fragmentation that described digestion products is three size respectively 294bp, 215bp and 79bp, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If the DNA fragmentation that described digestion products is three size respectively 294bp, 229bp and 78bp, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If the DNA fragmentation that described digestion products is four size respectively 215bp, 79bp, 229bp and 78bp, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
O, detect in Wheat volatiles DNA to be measured the sequence of DNA fragmentation of the 70th-240 corresponding to sequence 1, if corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O1), described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O2), described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O3), described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O1) and described O2), described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O1) and described O3), described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O2) and described O3), described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
Described O1) for the 70th-240 of sequence 1；
Described O2) for the 70th-240 of sequence 2；
Described O3) for the 70th-254 of sequence 3.
Wherein, the mass of 1000 kernel of A1A1 genotype wheat is higher than the mass of 1000 kernel of A3A3 genotype wheat, and the mass of 1000 kernel of A2A2 genotype wheat is higher than the mass of 1000 kernel of A3A3 genotype wheat；Every fringe spikelet number of A1A1 genotype wheat is less than every fringe spikelet number of A3A3 genotype wheat, and every fringe spikelet number of A2A2 genotype wheat is less than every fringe spikelet number of A3A3 genotype wheat；The single-strain tassel number of A1A1 genotype wheat is less than the single-strain tassel number of A3A3 genotype wheat.
Above-mentioned qualification or auxiliary are identified in the method for wheat genotypes, utilize the described primer pair P reaction system carrying out pcr amplification to contain: PCRbuffer, described p1 and described p2, dNTPs, archaeal dna polymerase, genomic DNA and water.Described reaction system is concretely: ddH2O8.0 μ L, 5 × PCRbuffer3.0 μ L, primer p1 (5 μm of ol/L) and p2 (5 μm of ol/L) each 0.6 μ L, dNTPs (2.5 μm of ol/L) 0.4 μ L, transfastpfu enzyme (5U) 0.3 μ L, genomic DNA (20ng/ μ L) 2.1 μ L.Wherein, 5 × PCRbuffer and transfastpfu (5U) can be all Beijing Quanshijin Biotechnology Co., Ltd's product, and dNTPs is Roche Products.
Described pcr amplification annealing temperature can be 59 DEG C.The condition of described pcr amplification is concretely: 95 DEG C of 5min, 95 DEG C of 30s, 59 DEG C of 30s, 72 DEG C of 30s, 35 circulations；72 DEG C of 10min.
Above-mentioned qualification or auxiliary are identified in the method for wheat genotypes, when detecting the sequence of DNA fragmentation of the 70th-240 corresponding to sequence 1 in Wheat volatiles DNA to be measured, as long as the sequence of this segment DNA can be detected, as detected by the method for DNA hybridization, specifically can hybridize such as the Southern marking.
For solving above-mentioned technical problem, present invention also offers identify or auxiliary identify wheat yield correlated traits method, described method is following 1), 2) or 3):
1) identify or assist the method identifying thousand grain weight of wheat character, including identifying that according to described qualification or auxiliary the method for wheat genotypes identifies the genotype of Semen Tritici aestivi to be measured, genotype according to Semen Tritici aestivi to be measured determines the thousand grain weight properties of described Semen Tritici aestivi to be measured: the mass of 1000 kernel of A1A1 genotype Semen Tritici aestivi to be measured higher than or candidate higher than the mass of 1000 kernel of A3A3 genotype Semen Tritici aestivi to be measured, the mass of 1000 kernel of A2A2 genotype Semen Tritici aestivi to be measured higher than or candidate higher than the mass of 1000 kernel of A3A3 genotype Semen Tritici aestivi to be measured；
2) identify or assist the method identifying per fringe spikelet number character, including identifying that according to described qualification or auxiliary the method for wheat genotypes identifies the genotype of Semen Tritici aestivi to be measured, genotype according to Semen Tritici aestivi to be measured determines every fringe spikelet number character of described Semen Tritici aestivi to be measured: every fringe spikelet number of A1A1 genotype Semen Tritici aestivi to be measured less than or candidate less than every fringe spikelet number of A3A3 genotype Semen Tritici aestivi to be measured, every fringe spikelet number of A2A2 genotype Semen Tritici aestivi to be measured less than or candidate less than every fringe spikelet number of A3A3 genotype Semen Tritici aestivi to be measured；
3) identify or assist the method identifying Semen Tritici aestivi single-strain tassel number character, including identifying that according to described qualification or auxiliary the method for wheat genotypes identifies the genotype of Semen Tritici aestivi to be measured, determine the single-strain tassel number character of described Semen Tritici aestivi to be measured according to the genotype of Semen Tritici aestivi to be measured: the single-strain tassel number of A1A1 genotype Semen Tritici aestivi to be measured less than or candidate less than the single-strain tassel number of A3A3 genotype Semen Tritici aestivi to be measured.
Above-mentioned qualification or auxiliary identify that in the method for wheat yield correlated traits, described Semen Tritici aestivi to be measured is not A1A3 genotype wheat, A1A2 genotype wheat and A2A3 genotype wheat.
For solving above-mentioned technical problem, present invention also offers CAPS-7A.
For solving above-mentioned technical problem, present invention also offers any one in following I-VI:
I, described primer pair P is in following Z1-Z5 application in any one；
Z1, qualification or auxiliary identify wheat genotypes；
Z2, preparation are identified or auxiliary qualification wheat genotypes product；
Z3, qualification or auxiliary identify wheat yield correlated traits；Described wheat yield correlated traits is thousand grain weight of wheat, every fringe spikelet number and/or single-strain tassel number；
Z4, preparation are identified or the auxiliary described wheat yield correlated traits product of qualification；
Z5, wheat breeding；
II, described reagent set is in above-mentioned Z1-Z5 application in any one；
III, described qualification or auxiliary identify the application in above-mentioned Z3 or Z5 of the method for wheat genotypes；
IV, described qualification or auxiliary identify the application in above-mentioned Z5 of the method for wheat yield correlated traits；
V, CAPS-7A application in above-mentioned Z5.
VI, the material of CAPS-7A is detected in above-mentioned Z1-Z5 application in any one.
Described detection CAPS-7A material can be by described reagent set with other reagent carried out needed for pcr amplification and/or instrument.Described other reagent carried out needed for pcr amplification can be containing dATP, dTTP, the dNTPs of dCTP and dGTP, archaeal dna polymerase and/or PCR reaction buffer；The described instrument carried out needed for pcr amplification can be PCR instrument.
For solving above-mentioned technical problem, present invention also offers method for breeding wheat, described method includes identifying that the method for wheat genotypes identifies the genotype of Semen Tritici aestivi according to described qualification or auxiliary, selects the genotypic Semen Tritici aestivi of A1A1, A2A2 or A3A3 to carry out breeding as parent.
In the present invention, described wheat yield correlated traits can be thousand grain weight of wheat, every fringe spikelet number and/or single-strain tassel number.
In the present invention, when detecting the size of PCR primer, electrophoresis detection can be passed through, it is possible to by detection of checking order.
Experiment proves, the present inventor is according to being found that a wheat molecular marker CAPS-7A relevant to wheat yield, according to CAPS-7A, Semen Tritici aestivi is carried out genotyping, three kinds can be obtained altogether and be homozygous genotype, be i.e. A1A1 genotype, A2A2 genotype and A3A3 genotype.Every fringe spikelet number all pole of A1A1 and A2A2 genotype wheat is substantially less than A3A3 genotype wheat, and the every fringe spikelet number between A1A1 and A2A2 genotype wheat is without significant difference；The single-strain tassel number pole of A1A1 genotype wheat is substantially less than A3A3 genotype wheat, and the single-strain tassel number between A1A1 and A2A2 genotype wheat is without significant difference, and the single-strain tassel number between A2A2 and A3A3 genotype wheat is without significant difference；The mass of 1000 kernel of A1A1 and A2A2 genotype wheat is all remarkably higher than A3A3 genotype wheat, and the mass of 1000 kernel between A1A1 and A2A2 genotype wheat is without significant difference.Illustrate that the CAPS-7A in the present invention can have in excellent yield traits at selection-breeding Semen Tritici aestivi to work.The present invention is that wheat molecular marker assisted selection provides a new method, significant in cultivating high yield wheat breed or research.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention being further described in detail, the embodiment provided is only for illustrating the present invention, rather than in order to limit the scope of the present invention.
Experimental technique in following embodiment, if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all commercially obtain.
5 × PCRbuffer and transfastpfu (5U) in following embodiment is Beijing Quanshijin Biotechnology Co., Ltd's product, and catalog number is AP221-02；DNTPs is Roche Products, and product article No. is #316K5S.
Nco I in following embodiment is Fermentas Products, and catalog number is ER0575.
Embodiment 1, CAPS-7A are relevant to wheat yield correlated traits
One, the discovery of CAPS-7A
The present inventor (comes from country's germplasm resource bank according to 32 parts of Guard cell kinds in table 1, the public can obtain from country's germplasm resource bank) it is found that a wheat molecular marker relevant to wheat yield, by its called after CAPS-7A, this CAPS-7A is template with Wheat volatiles DNA, utilizes primer pair P to carry out the PCR primer that pcr amplification obtains, different wheat breeds has three kinds of sequences, sequence 1, sequence 2 and sequence 3.By name, primer pair P is called that the single stranded DNA of p1 and p2 forms, p1 is the single stranded DNA shown in sequence 4 (5 '-AGTCTATACCCCACACCCCATC-3 '), and p2 is the single stranded DNA shown in sequence 5 (5 '-GCATTGAGTAGTGTTTTGTGCTGT-3 ').
By the Semen Tritici aestivi called after A1A1 genotype wheat that PCR primer sequence is sequence 1；By the Semen Tritici aestivi called after A2A2 genotype wheat that PCR primer sequence is sequence 2；By the Semen Tritici aestivi called after A3A3 genotype wheat that PCR primer sequence is sequence 3；By the Semen Tritici aestivi called after A1A2 genotype wheat that PCR primer sequence is sequence 1 and sequence 2；By the Semen Tritici aestivi called after A1A3 genotype wheat that PCR primer sequence is sequence 1 and sequence 3；By the Semen Tritici aestivi called after A2A3 genotype wheat that PCR primer sequence is sequence 2 and sequence 3.
Wherein, sequence 2 and the DNA molecular shown in sequence 3 all contain the recognition sequence of restricted enzyme Nco I, with Nco I, the DNA molecular shown in sequence 2 is carried out enzyme action, the DNA fragmentation of two size respectively 215bp and 79bp can be obtained, with Nco I, the DNA molecular shown in sequence 3 is carried out enzyme action, it is possible to obtain the DNA fragmentation of two size respectively 229bp and 78bp.
1,32 parts of Guard cell kinds of table
| Numbering | Title | Numbering | Title |
| 1 | PANDAS | 17 | Face anti-5108 |
| 2 | 124-1 in peace 85 | 18 | Bai Qimai |
| 3 | Lay down and open up No. one | 19 | Changle 5 |
| 4 | Euphorbia royleana Boiss. | 20 | Red Buddhist monk |
| 5 | Beijing 10 | 21 | Beijing 8686 |
| 6 | Beijing 14 | 22 | 04-044 |
| 7 | Cangzhou Semen Tritici aestivi | 23 | 04-030 |
| 8 | Changwu 131 | 24 | Spring 22 9th-25 |
| 9 | Long 6878 | 25 | Purple stalk Bai Mangxian |
| 10 | Dali 1 | 26 | Capital product 10 |
| 11 | Single R8093 | 27 | Spring 04 9th-5-1 |
| 12 | Rich anti-13 | 28 | Spring 45 9th-50-1 |
| 13 | Ji wheat 41 | 29 | Interior township 188 |
| 14 | Ji wheat No. 6 | 30 | Capital 411 |
| 15 | Shanxi 2148-7 | 31 | China spring |
| 16 | Capital core 8922 | 32 | White rough wheat |
Two, CAPS-7A is relevant to wheat yield correlated traits
Choose 262 parts of hexaploid wheats composition natural population 1 (table 2), adopt the CAPS-7A labelling in step one that natural population 1 carries out genotyping, and be associated analyzing to genotype and every fringe spikelet number, single-strain tassel number and three Correlated Yield Characters of mass of 1000 kernel.
1, genotypic determine
1) extract the genomic DNA of Semen Tritici aestivi to be measured, carry out pcr amplification with genome specific primer (F and R), obtain pcr amplification product A；
F:5 '-TCCAAAACAACCACGGCTAAC-3 '；
R:5′-AGGACTCGGCCAAGAATACAAG-3′。
2) with dilute 150 times pcr amplification product A for template, adopt the primer pair P in step one to carry out pcr amplification, obtain pcr amplification product B；
The system (15 μ L) of pcr amplification is: ddH2O8.0 μ L, 5 × PCRbuffer3.0 μ L, primer p1 (5 μm of ol/L) and p2 (5 μm of ol/L) each 0.6 μ L, dNTPs (2.5 μm of ol/L) 0.4 μ L, transfastpfu enzyme (5U) 0.3 μ L, genomic DNA (20ng/ μ L) 2.1 μ L；
Pcr amplification condition is: 95 DEG C of 5min, 95 DEG C of 30s, 59 DEG C of 30s, 72 DEG C of 30s, 35 circulations；72 DEG C of 10min, 4 DEG C of preservations.
3) pcr amplification product B being checked order, according to the method for genotyping in step one, each plant in natural population 1 is carried out genotyping, result is as shown in table 2.
Each Semen Tritici aestivi haplotype statistical result of table 2, natural population 1
Note: "-" indicates without PCR primer.
4) check order with restricted enzyme Nco I enzyme action pcr amplification product B, according to the method in step one, Semen Tritici aestivi carried out genotyping, result and step 3 according to the size of digestion products) in genotyping result consistent.
2, the association analysis of genotype and Correlated Yield Characters
Every fringe spikelet number of each wheat breed, single-strain tassel number and mass of 1000 kernel is investigated during plantation.GLM model is utilized to be associated three kinds of haplotypes and correlated traits analyzing with Tassel2.1 software.
It was found that every fringe spikelet number all pole of A1A1 and A2A2 genotype wheat is substantially less than A3A3 genotype wheat, the every fringe spikelet number between A1A1 and A2A2 genotype wheat is without significant difference；The single-strain tassel number pole of A1A1 genotype wheat is substantially less than A3A3 genotype wheat, A1A1 and A3A3 genotype respectively with the single-strain tassel number of A2A2 genotype wheat without significant difference；The mass of 1000 kernel of A1A1 and A2A2 genotype wheat is all remarkably higher than A3A3 genotype wheat, and the mass of 1000 kernel between A1A1 and A2A2 genotype wheat is without significant difference.Illustrate that the CAPS-7A in the present invention can have in excellent yield traits at selection-breeding Semen Tritici aestivi to work.
Three kinds of genotype Correlated Yield Characters statistical result in table 3, Semen Tritici aestivi natural population 1
| Character | A1A1 | A2A2 | A3A3 |
| Every fringe spikelet number (individual) | 18.43±0.12B | 18.24±0.21B | 18.92±0.11A |
| Single-strain tassel number (individual) | 12.96±0.25B | 13.58±0.54AB | 14.03±0.32A |
| Mass of 1000 kernel (g) | 42.59±0.47a | 42.54±0.84a | 40.79±0.55b |
Note: lower case and upper case letter represents Traits change notable (P < 0.05) and extremely notable (P < 0.01) between different genetic wheat varieties respectively.
Claims (10)
1. wheat molecular marker is in the application identified or in auxiliary qualification wheat yield correlated traits；
Described wheat molecular marker, for being template with the genomic DNA of Semen Tritici aestivi, adopt primer pair P to carry out expanding the DNA molecular obtained；By name, described primer pair P is called that the single stranded DNA of p1 and p2 forms, described p1 is and the single stranded DNA of the 70th upstream specific bond of sequence 1 in Wheat volatiles DNA, and described p2 is and the single stranded DNA of the 241st downstream specific bond of sequence 1 in Wheat volatiles DNA.
2. application according to claim 1, it is characterised in that: described p1 is the single stranded DNA shown in sequence 4, and described p2 is the single stranded DNA shown in sequence 5.
3. identify or assist the primer pair identifying wheat yield correlated traits, for primer pair P described in claim 1 or 2.
4. identifying or assist the reagent set identifying wheat yield correlated traits, described in claim 1 or 2, primer pair P and restricted enzyme Nco I forms.
5. application according to claim 1 and 2, or the primer pair described in claim 3, or the reagent set described in claim 4, it is characterised in that: described wheat yield correlated traits is thousand grain weight of wheat, every fringe spikelet number and/or single-strain tassel number.
6. identifying or assist the method identifying wheat genotypes, described genotype is A1A1 genotype, A2A2 genotype, A3A3 genotype, A1A2 genotype, A1A3 genotype and A2A3 genotype, and described method is following L, M, N or O:
L, include following L1) and L2):
L1) with Wheat volatiles DNA to be measured for template, utilize primer pair P described in claim 1 or 2 to carry out pcr amplification and obtain PCR primer；
L2) detecting step L1) sequence of PCR primer that obtains, if described PCR primer contains the DNA fragmentation shown in X1 but do not contain the DNA fragmentation shown in X2 or X3, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X2 and without there being the DNA fragmentation shown in described X1 or described X3, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X3 and without there being the DNA fragmentation shown in described X1 or described X2, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X1 and described X2 and without there being the DNA fragmentation shown in described X3, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X1 and described X3 and without there being the DNA fragmentation shown in described X2, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If described PCR primer contains the DNA fragmentation shown in described X2 and described X3 and without there being the DNA fragmentation shown in described X1, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
Described X1 is the 70th-240 of sequence 1；
Described X2 is the 70th-240 of sequence 2；
Described X3 is the 70th-254 of sequence 3；
M, include following M1) and M2):
M1) with Wheat volatiles DNA to be measured for template, utilize primer pair P described in claim 2 to carry out pcr amplification and obtain PCR primer；
M2) detecting step M1) sequence of PCR primer that obtains, if the sequence of described PCR primer is sequence 1, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If the sequence of described PCR primer is sequence 2, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If the sequence of described PCR primer is sequence 3, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If the sequence of described PCR primer is sequence 1 and sequence 2, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If the sequence of described PCR primer is sequence 1 and sequence 3, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If the sequence of described PCR primer is sequence 2 and sequence 3, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
N, include following N1) and N2):
N1) with Wheat volatiles DNA to be measured for template, utilize primer pair P described in claim 2 to carry out pcr amplification and obtain PCR primer, adopt restricted enzyme NcoI to process described PCR primer and obtain digestion products；
N2) detecting step N1) size of digestion products that obtains, if the DNA fragmentation that described digestion products is a 294bp, described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If the DNA fragmentation that described digestion products is two size respectively 215bp and 79bp, described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If the DNA fragmentation that described digestion products is two size respectively 229bp and 78bp, described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If the DNA fragmentation that described digestion products is three size respectively 294bp, 215bp and 79bp, described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If the DNA fragmentation that described digestion products is three size respectively 294bp, 229bp and 78bp, described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If the DNA fragmentation that described digestion products is four size respectively 215bp, 79bp, 229bp and 78bp, described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
O, detect in Wheat volatiles DNA to be measured the sequence of DNA fragmentation of the 70th-240 corresponding to sequence 1, if corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O1), described Semen Tritici aestivi to be measured is A1A1 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O2), described Semen Tritici aestivi to be measured is A2A2 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is O3), described Semen Tritici aestivi to be measured is A3A3 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O1) and described O2), described Semen Tritici aestivi to be measured is A1A2 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O1) and described O3), described Semen Tritici aestivi to be measured is A1A3 genotype wheat；If corresponding to the sequence of the DNA fragmentation of the 70th of sequence 1 the-240 is described O2) and described O3), described Semen Tritici aestivi to be measured is A2A3 genotype wheat；
Described O1) for the 70th-240 of sequence 1；
Described O2) for the 70th-240 of sequence 2；
Described O3) for the 70th-254 of sequence 3.
7. identify or auxiliary identify wheat yield correlated traits method, for following 1), 2) or 3):
1) identify or assist the method identifying thousand grain weight of wheat character, the genotype of Semen Tritici aestivi to be measured is identified including method described in claim 6, genotype according to Semen Tritici aestivi to be measured determines the thousand grain weight properties of described Semen Tritici aestivi to be measured: the mass of 1000 kernel of A1A1 genotype Semen Tritici aestivi to be measured higher than or candidate higher than the mass of 1000 kernel of A3A3 genotype Semen Tritici aestivi to be measured, the mass of 1000 kernel of A2A2 genotype Semen Tritici aestivi to be measured higher than or candidate higher than the mass of 1000 kernel of A3A3 genotype Semen Tritici aestivi to be measured；
2) identify or assist the method identifying per fringe spikelet number character, the genotype of Semen Tritici aestivi to be measured is identified including method described in claim 6, genotype according to Semen Tritici aestivi to be measured determines every fringe spikelet number character of described Semen Tritici aestivi to be measured: every fringe spikelet number of A1A1 genotype Semen Tritici aestivi to be measured less than or candidate less than every fringe spikelet number of A3A3 genotype Semen Tritici aestivi to be measured, every fringe spikelet number of A2A2 genotype Semen Tritici aestivi to be measured less than or candidate less than every fringe spikelet number of A3A3 genotype Semen Tritici aestivi to be measured；
3) identify or assist the method identifying Semen Tritici aestivi single-strain tassel number character, identify the genotype of Semen Tritici aestivi to be measured including method described in claim 6, determine the single-strain tassel number character of described Semen Tritici aestivi to be measured according to the genotype of Semen Tritici aestivi to be measured: the single-strain tassel number of A1A1 genotype Semen Tritici aestivi to be measured less than or candidate less than the single-strain tassel number of A3A3 genotype Semen Tritici aestivi to be measured.
8. wheat molecular marker described in claim 1.
9. any one in following I-VI:
I, primer pair described in claim 3 or 5 is in following Z1-Z5 application in any one；
Z1, qualification or auxiliary identify wheat genotypes；
Z2, preparation are identified or auxiliary qualification wheat genotypes product；
Z3, qualification or auxiliary identify wheat yield correlated traits；Described wheat yield correlated traits is thousand grain weight of wheat, every fringe spikelet number and/or single-strain tassel number；
Z4, preparation are identified or the auxiliary described wheat yield correlated traits product of qualification；
Z5, wheat breeding；
II, reagent set described in claim 4 or 5 is in above-mentioned Z1-Z5 application in any one；
III, identify described in claim 6 or auxiliary identifies the application in above-mentioned Z3 or Z5 of the method for wheat genotypes；
IV, identify described in claim 7 or auxiliary identifies the application in above-mentioned Z5 of the method for wheat yield correlated traits；
V, the application in above-mentioned Z5 of the wheat molecular marker described in claim 1；
VI, test right requires that the material of wheat molecular marker described in 1 is in above-mentioned Z1-Z5 application in any one.
10. method for breeding wheat, identifies the genotype of Semen Tritici aestivi in accordance with the method for claim 6, selects A1A1 genotype, A2A2 genotype or the genotypic Semen Tritici aestivi of A3A3 to carry out breeding as parent.
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